U.S. patent number 7,297,693 [Application Number 10/488,579] was granted by the patent office on 2007-11-20 for nitrogen-containing heterocyclic compounds and their use as raf inhibitors.
This patent grant is currently assigned to SmithKline Beecham P.L.C.. Invention is credited to Mark James Bamford, David Kenneth Dean, Antoinette Naylor, Andrew Kenneth Takle, David Matthew Wilson.
United States Patent |
7,297,693 |
Bamford , et al. |
November 20, 2007 |
Nitrogen-containing heterocyclic compounds and their use as Raf
inhibitors
Abstract
Compounds and their use as pharmaceuticals particularly as Raf
kinase inhibitors for the treatment of neurotraumatic diseases,
cancer, chronic neurodegeneration, pain, migraine and cardiac
hypertrophy.
Inventors: |
Bamford; Mark James (Harlow,
GB), Dean; David Kenneth (Harlow, GB),
Naylor; Antoinette (Harlow, GB), Takle; Andrew
Kenneth (Harlow, GB), Wilson; David Matthew
(Harlow, GB) |
Assignee: |
SmithKline Beecham P.L.C.
(Middlesex, GB)
|
Family
ID: |
9921562 |
Appl.
No.: |
10/488,579 |
Filed: |
September 5, 2002 |
PCT
Filed: |
September 05, 2002 |
PCT No.: |
PCT/EP02/09942 |
371(c)(1),(2),(4) Date: |
March 04, 2004 |
PCT
Pub. No.: |
WO03/022836 |
PCT
Pub. Date: |
March 20, 2003 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20040209883 A1 |
Oct 21, 2004 |
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Foreign Application Priority Data
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Sep 5, 2001 [GB] |
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0121494.9 |
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Current U.S.
Class: |
514/235.5;
514/304; 514/299; 514/253.1; 514/316; 514/336; 514/343; 546/112;
546/125; 546/187; 546/193; 546/279.1; 546/284.7; 514/318 |
Current CPC
Class: |
A61P
43/00 (20180101); C07D 451/04 (20130101); C07D
405/14 (20130101); A61P 25/04 (20180101); A61P
25/28 (20180101); C07D 409/04 (20130101); A61P
25/06 (20180101); C07D 405/04 (20130101); A61P
9/00 (20180101); A61P 35/00 (20180101); A61P
25/00 (20180101); A61P 25/16 (20180101); A61P
9/10 (20180101) |
Current International
Class: |
A61K
31/506 (20060101); A61K 31/443 (20060101); A61K
31/4439 (20060101); C07D 401/04 (20060101); C07D
405/04 (20060101); C07D 405/14 (20060101); C07D
407/14 (20060101) |
Field of
Search: |
;544/121,131
;546/112,125,187,193,279.1,284.7
;514/235.5,253.1,299,304,316,318,336,343 |
Foreign Patent Documents
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WO 00/25791 |
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May 2000 |
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WO |
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WO 01/66540 |
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Sep 2001 |
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WO |
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Other References
Wermuth, C.G. (ed.), "Chapter 13: Molecular Variations Based on
Isosteric Replacements", The Practice of Medicinal Chemistry, pp.
204-237, Academic Press Ltd., Copyright (1996), XP-002190259. cited
by other.
|
Primary Examiner: Dentz; Bernard
Attorney, Agent or Firm: Hsu; Grace C. McCarthy; Mary E.
Kinzig; Charles M.
Claims
The invention claimed is:
1. A compound of formula (I): ##STR00013## wherein: X is O,
CH.sub.2, CO, S or NH, or the moiety X--R.sup.1 is hydrogen;
Y.sub.1 and Y.sub.2 independently represent CH or N; R.sup.1 is
hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl, aryl,
arylC.sub.1-6alkyl-, heterocyclyl, heterocyclylC.sub.1-6alkyl-,
heteroaryl, or heteroarylC.sub.1-6alkyl-, any of which, except
hydrogen, may be optionally substituted; R.sup.2 is
CONR.sup.6R.sup.7; R.sup.6 and R.sup.7 independently represent
hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl, aryl,
arylC.sub.1-6alkyl, heteroaryl, heteroarylC.sub.1-6alkyl,
heterocyclyl, or heterocyclylC.sub.1-6alkyl, any of which except
for hydrogen may be optionally substituted, or R.sup.6 and R.sup.7
together with the nitrogen atom to which they are attached form a
3- to 12-membered monocyclic or bicyclic ring optionally including
up to three heteroatoms selected from O, N or S wherein said ring
may be optionally substituted; Ar is a group of the formula a) or
b): ##STR00014## wherein A represents a fused 5- to 7-membered ring
optionally containing up to two heteroatoms selected from O, S and
NR.sup.5, wherein R.sup.5 is hydrogen or C.sub.1-6alkyl, which ring
is optionally substituted by up to 2 substituents selected from
halogen, C.sub.1-6alkyl, hydroxy, C.sub.1-6alkoxy or keto; R.sup.3
and R.sup.4 are independently selected from hydrogen, halogen,
C.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxyC.sub.1-6alkyl, haloC.sub.1-6alkyl,
arylC.sub.1-6alkoxy, hydroxy, nitro, cyano, azido, amino, mono- and
di-N-C.sub.1-6alkylamino, acylamino, arylcarbonylamino, acyloxy,
carboxy, carboxy salts, carboxy esters, carbamoyl, mono- and
di-N-C.sub.1-6alkylcarbamoyl, C.sub.1-6alkoxycarbonyl,
aryloxycarbonyl, ureido, guanidino, C.sub.1-6alkylguanidino,
amidino, C.sub.1-6alkylamidino, sulphonylamino, aminosulphonyl,
C.sub.1-6alkylthio, C.sub.1-6alkyl sulphinyl or
C.sub.1-6alkylsulphonyl; and one of X.sub.1 and X.sub.2 is selected
from O, S or NR.sup.11 and the other is CH, wherein R.sup.11 is
hydrogen, C.sub.1-6alkyl, aryl or arylC.sub.1-6alkyl; or a
pharmaceutically acceptable salt thereof.
2. The compound according to claim 1, wherein X--R.sup.1 is
hydrogen.
3. The compound according to claim 1, wherein A represents a fused
5 membered ring optionally containing up to two heteroatoms
selected from O, S and NR.sup.5, wherein R.sup.5 is hydrogen or
C.sub.1-6alkyl, which ring is optionally substituted by up to 2
substituents selected from halogen, C.sub.1-6alkyl, hydroxy,
C.sub.1-6alkoxy or keto.
4. The compound according to claim 1, wherein R.sup.6 and R.sup.7
are independently selected from hydrogen, C.sub.1-6alkyl, aryl,
heterocyclic and heterocyclicC.sub.1-6alkyl, wherein any of the
groups except hydrogen may be optionally substituted or R.sup.6 and
R.sup.7 together with the nitrogen atom to which they are attached
form a 3- to 7-membered monocyclic optionally including up to three
heteroatoms selected from O, N or S wherein said ring may be
optionally substituted.
5. A compound, wherein the compound is:
5-[5-(1-Morpholin-4-yl-methanoyl)-2-pyridin-4-yl-furan-3-yl]-indan-1-one
oxime;
5-[5-(1-Piperidin-1-yl-methanoyl)-2-pyridin-4-yl-furan-3-yl]-indan-
-1-one oxime;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid (2-morpholin-4-yl-ethyl)-amide;
5-(5-{1-[4-(2-Methoxy-ethyl)-piperazin-1-yl]-methanoyl}-2-pyridin-4-yl-fu-
ran-3-yl)-indan-1-one oxime;
4-(1-Hydroximino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic acid
methyl-(1-methyl-piperidin-4-yl)-amide;
5-[5-(1-[1,4']Bipiperidinyl-1'-yl-methanoyl)-2-pyridin-4-yl-furan-3-yl]-i-
ndan-1-one oxime;
5-(5-{1-[4-(4-Chloro-benzyl)-piperazin-1-yl]-methanoyl}-2-pyridin-4-yl-fu-
ran-3-yl)-indan-1-one oxime;
5-{5-[1-(4-Cyclohexyl-piperazin-1-yl)-methanoyl]-2-pyridin-4-yl-furan-3-y-
l}-indan-1one oxime;
5-{5-[1-(4-Methyl-piperazin-1-yl)-methanoyl]-2-pyridin-4-yl-fu
ran-3-yl}-indan-1-one oxime;
5-(5-{1-[4-(2-Oxo-2-pyrrolidin-1-yl-ethyl)-piperazin-1-yl]methanoyl}-2-py-
ridin-4-yl-furan-3-yl)-indan-1-one oxime;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid 4-dimethyamino-benzylamide;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid (2-dimethylaminoethyl)-methylamide;
4-[({1-[4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-yl]-methanoy-
l}-amino)-methyl]-piperidine-1-carboxylic acid tert-butyl ester;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid (2-dimethylamino-ethyl)-amide;
5-(2-Pyridin-4-yl-5-{1-[4-(2-pyrrolidin-1-yl-ethyl)-piperazin-1-yl]-metha-
noyl}-furan-3-yl)-indan-1-one oxime;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid (1-piperidin-4-yl-cyclohexylmethyl)-amide;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid [4-(2-dimethylamino-ethoxy)-phenyl]-amide;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid (2-pyrrolidin-1-yl-ethyl)-amide;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid [3-(4-methyl-piperazin-1-yl)-propyl]-amide;
5-{2-Pyridin-4-yl-5-[1-(4-pyrrolidin-1-yl-piperidin-1-yl]-methanoyl}-fura-
n-3-yl}-indan-1-one oxime;
5-(5-{1-[4-(2-Dimethylamino-ethyl)-piperazin-1-yl]-methanoyl}-2-pyridin-4-
-yl-furan-3-yl)-indan-1-one oxime;
5-(5-{1-[4-(2-Dimethylamino-ethyl)-piperazin-1-yl]-methanoyl}-2-pyridin-4-
-yl-furan-3-yl)-indan-1-one oxime;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid (2-diisopropyl amino-ethyl)-amide;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid ((1R,5S) -8-methyl-azabicyclo[3.2.1]oct-3-yl)amide;
4-(1-Hydroximino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic acid
((1R,5S)-9-methyl-9-azabicyclo[3.3.1]non-3-yl)amide;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid methyl-piperidin-4-yl-amide;
5-[5-(1-Piperazin-1-yl-methanoyl)-2-pyridin-4-yl)-furan-3-yl]-indan-1-one
oxime;
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid methyl-piperidin-4-yl-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-dimethylamino-ethyl)-methyl-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid methyl-(1-methyl -piperidin-4-yl)-amide;
5-[5-(1-[1,4']Bipiperidinyl-1'-yl-methanoyl)-3-pyridin-4-yl-furan-2-yl]-i-
ndan-1-one oxime;
5-(5-{1-[4-(4-Chloro-benzyl)-piperazin-1-yl]-methanoyl}-3-pyridin-4-yl-fu-
ran-2-yl)-indan-1-one oxime;
5-{5-[1-(4-Cyclohexyl-piperazin-1-yl)-methanoyl]-3-pyridin-4-yl-furan-2-y-
l}-indan-1-one oxime;
5-{5-[1-(4-Methyl-piperazin-1-yl)-methanoyl]-3-pyridin-4-yl-furan-2-yl}-i-
ndan-1-one oxime;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-pyrrolidin-1-yl-ethyl)-amide;
5-(5-{1-[4-(2-Oxo-2-pyrrolidin-1-yl-ethyl)-piperazin-1-yl]-methanoyl}-3-p-
yridin-4-yl-furan-2-yl)-indan-1-one oxime;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid 4-dimethylamino-benzylamide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-diisopropylamino-ethyl)-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-diisopropylamino-ethyl)-amide;
4-[({1-[5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-yl]-methanoy-
l}-amino)-methyl]-piperidine-1-carboxylic acid tert-butyl ester;
5-(3-Pyridin-4-yl-5-{1-[4-(2-pyrrolidin-1-yl-ethyl)-piperazin-1-yl]-metha-
noyl}-furan-2-yl)-indan-1-one oxime;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-5hydroxy-ethyl)-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-pyrrolidin-1-yl-ethyl)-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-dimethylamino-ethyl)-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid [3-(4-methyl-piperazin-1-yl)-propyl]-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (1-piperidin-1-yl-cyclohexylmethyl)-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid [4-(2-dimethylamino-ethoxy)-phenyl]-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (3-dimethylamino-propyl)-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-piperidin-1-yl-ethyl)-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-amino-ethyl)-amide;
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid piperidin-4-ylamide;
(+/-)5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid pyrrolidin-3-ylamide; and
5-(1-Hydroximino-indan-5-yl)-4-pyridin-4-yl-thiophene-2-carboxylic
acid methyl-(1-methyl-piperidin-4-yl)-amide.
6. A pharmaceutical composition comprising a compound according to
claim 1 or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
7. A method for therapeutic treatment of colorectal cancer in a
human, or other mammal, which comprises administering a
therapeutically effective amount of a compound of claim one of
formula (I) or a pharmaceutically acceptable salt thereof.
8. A method for therapeutic treatment of melanoma in a human, or
other mammal, which comprises administering a therapeutically
effective amount of a compound of claim one of formula (I) or a
pharmaceutically acceptable salt thereof.
Description
This application is a .sctn.371 national stage filing of
PCT/EP02/09942 filed 5 Sep. 2002.
This invention relates to novel compounds and their use as
pharmaceuticals, particularly as Raf kinase inhibitors for the
treatment of neurotraumatic diseases, cancer chronic
neurodegeneration, pain, migraine and cardiac hypertrophy.
Raf protein kinases are key components of signal transduction
pathways by which specific extracellular stimuli elicit precise
cellular responses in mammalian cells. Activated cell surface
receptors activate ras/rap proteins at the inner aspect of the
plasma-membrane which in turn recruit and activate Raf proteins.
Activated Raf proteins phosphorylate and activate the intracellular
protein kinases MEK1 and MEK2. In turn, activated MEKs catalyse
phosphorylation and activation of p42/p44 mitogen-activated protein
kinase (MAPK). A variety of cytoplasmic and nuclear substrates of
activated MAPK are known which directly or indirectly contribute to
the cellular response to environmental change. Three distinct genes
have been identified in mammals that encode Raf proteins; A-Raf,
B-Raf and C-Raf (also known as Raf-1) and isoformic variants that
result from differential splicing of mRNA are known.
Inhibitors of Raf kinases have been suggested for use in disruption
of tumor cell growth and hence in the treatment of cancers, e.g.
histiocytic lymphoma, lung adenocarcinoma, small cell lung cancer
and pancreatic and breast carcinoma; also in the treatment and/or
prophylaxis of disorders associated with neuronal degeneration
resulting from ischemic events, including cerebral ischemia after
cardiac arrest, stroke and multi-infarct dementia and also after
cerebral ischemic events such as those resulting from head injury,
surgery and/or during childbirth; also in chronic neurodegeneration
such as Alzheimer's disease and Parkinson's disease; also in the
treatment of pain, migraine and cardiac hypertrophy.
We have now found a group of novel compounds that are inhibitors of
Raf kinases, in particular inhibitors of B-Raf kinase.
According to the invention there is provided compounds of formula
(I):
##STR00001## wherein;
X is O, CH.sub.2, CO, S or NH, or the moiety X--R.sup.1 is
hydrogen;
Y.sub.1 and Y.sub.2 independently represent CH or N;
R.sup.1 is hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl, aryl,
arylC.sub.1-6alkyl-, heterocyclyl, heterocyclylC.sub.1-6alkyl-,
heteroaryl, or heteroarylC.sub.1-6alkyl-, any of which, except
hydrogen, may be optionally substituted;
R.sup.2 is CONR.sup.6R.sup.7;
R.sup.6 and R.sup.7 independently represent hydrogen,
C.sub.1-6alkyl, C.sub.3-7cycloalkyl, aryl, arylC.sub.1-6alkyl,
heteroaryl, heteroarylC.sub.1-6alkyl, heterocyclyl, or
heterocyclylC.sub.1-6alkyl, any of which except for hydrogen may be
optionally substituted, or R.sup.6 and R.sup.7 together with the
nitrogen atom to which they are attached form a 3- to 12-membered
monocyclic or bicyclic ring optionally including upto three
heteroatoms selected from O, N or S wherein said ring may be
optionally substituted;
Ar is a group of the formula a) or b):
##STR00002##
wherein A represents a fused 5- to 7-membered ring optionally
containing up to two heteroatoms selected from O, S and NR.sup.5,
wherein R.sup.5 is hydrogen or C.sub.1-6alkyl, which ring is
optionally substituted by up to 2 substituents selected from
halogen, C.sub.1-6alkyl, hydroxy, C.sub.1-6alkoxy or keto;
R.sup.3 and R.sup.4 are independently selected from hydrogen,
halogen, C.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxyC.sub.1-6alkyl, haloC.sub.1-6alkyl,
arylC.sub.1-6alkoxy, hydroxy, nitro, cyano, azido, amino, mono- and
di-N-C.sub.1-6alkylamino, acylamino, arylcarbonylamino, acyloxy,
carboxy, carboxy salts, carboxy esters, carbamoyl, mono- and
di-N-C.sub.1-6alkylcarbamoyl, C.sub.1-6alkoxycarbonyl,
aryloxycarbonyl, ureido, guanidino, C.sub.1-6alkylguanidino,
amidino, C.sub.1-6alkylamidino, sulphonylamino, aminosulphonyl,
C.sub.1-6alkylthio, C.sub.1-6alkyl sulphinyl or
C.sub.1-6alkylsulphonyl; and
one of X.sub.1 and X.sub.2 is selected from O, S or NR.sup.11 and
the other is CH, wherein R.sup.11 is hydrogen, C.sub.1-6alkyl, aryl
or arylC.sub.1-6alkyl;
or pharmaceutically acceptable salts thereof.
As used herein, the double bond indicated by the dotted lines of
formula (I), represents the possible regioisomeric ring forms of
the compounds falling within the scope of this invention, the
double bond being between the non-heteroatoms.
The hydroxyimino moiety can be positioned on any of carbon atoms of
the non-aromatic ring in groups a) and b).
The hydroxyimino moiety can exist as either the E or Z isomer or as
a mixture of both. Alkyl and alkenyl groups referred to herein,
individually or as part of larger groups e.g. alkoxy, may be
straight or branched groups containing up to six carbon atoms and
are optionally substituted by one or more groups selected from the
group consisting of aryl, heteroaryl, heterocyclyl,
C.sub.1-6alkoxy, C.sub.1-6alkylthio, arylC.sub.1-6alkoxy,
arylC.sub.1-6alkylthio, amino, mono- or di-C.sub.1-6alkylamino,
cycloalkyl, cycloalkenyl, carboxy and esters thereof, amide,
sulphonamido, ureido, guanidino, C.sub.1-6alkylguanidino, amidino,
C.sub.1-6alkylamidino, C.sub.1-6acyloxy, azido, hydroxy,
hydroxyimino and halogen.
Cycloalkyl and cycloalkenyl groups referred to herein include
groups having from three to seven ring carbon atoms and are
optionally substituted as described hereinabove for alkyl and
alkenyl groups.
Optional substituents for alkyl, alkenyl, cycloalkyl and
cycloalkenyl groups include aryl, heteroaryl, heterocyclyl,
C.sub.1-6alkoxy, C.sub.1-6alkylthio, arylC.sub.1-6alkoxy,
arylC.sub.1-6alkylthio, amino, mono- or di-C.sub.1-6alkylamino,
aminosulphonyl, cycloalkyl, cycloalkenyl, carboxy and esters
thereof, amide, ureido, quanidino, C.sub.1-6alkylquanidino,
amidino, C.sub.1-6alkylamidino, C.sub.1-6acyloxy, hydroxy, and
halogen or any combination thereof. Preferably the substituents are
mono-or di-C.sub.1-6alkylamino, heterocycloC.sub.1-6alkylamino or
C.sub.2-6acylamino.
Alternatively the optional substituent contains a
water-solubilising group; suitable solubilising moieties will be
apparent to those skilled in the art and include hydroxy and amine
groups. Even more preferably the optional substituent includes
amino, mono- or di-C.sub.1-6alkylamino, amine containing
heterocyclyl, or hydroxy or any combination thereof.
When used herein, the term "aryl" includes, unless otherwise
defined, single and fused rings suitably containing from 4 to 7,
preferably 5 or 6, ring atoms in each ring, which rings, may each
be unsubstituted or substituted by, for example, up to three
substituents.
Suitable aryl groups include phenyl and naphthyl, such as
1-naphthyl or 2-naphthyl.
When used herein in respect of R.sup.6 or R.sup.7, the term
"monocyclic ring" means a 3 to 7 membered ring system for example
phenyl, pyrrole, pyrroline, pyrrolidine, piperidine, morpholine,
thiomorpholine, piperizine, indole or indoline. The term "bicyclic
ring" means a 7 to 12 membered fused ring system e.g napthyl.
When used herein heteroC.sub.1-6alkyl- means a C.sub.1-6 carbon
chain wherein the end carbon atom in the chain is substituted by a
heteroatom selected from N, O, or S for example
C.sub.1-6alkylamino, C.sub.1-6alkyloxy or C.sub.1-6alkylthio.
C.sub.1-6alkylheteroC.sub.1-6alkyl means a C.sub.3-13alkyl chain
wherein one of the carbon atoms has been replaced with a heteroatom
selected from N, O, or S, for example
C.sub.1-6alkylaminoC.sub.1-6alkyl or C.sub.1-6
alkylaminodiC.sub.1-6alkyl, C.sub.1-6alkyloxyC.sub.1-6alkyl-,
C.sub.1-6alkylthioC.sub.1-6alkyl-, or
C.sub.1-6alkylthiodiC.sub.1-6alkyl.
When used herein the term "heterocyclyl" includes, unless otherwise
defined, non-aromatic, single or fused, saturated or unsaturated,
rings suitably containing up to four heteroatoms in each ring, each
of which is selected from O, N and S, which rings, may be
unsubstituted or substituted by, for example, up to three
substituents. Each heterocyclic ring suitably has from 4 to 7,
preferably 5 or 6, ring atoms. A fused heterocyclic ring system may
include carbocyclic rings and need include only one heterocyclic
ring. Examples of heterocyclyl groups include pyrrolidine,
piperidine, piperazine, morpholine, thiomorpholine, imidazolidine
and pyrazolidine.
When used herein, the term "heteroaryl" includes, unless otherwise
defined, mono- and bicyclic heteroaromatic ring systems comprising
up to four, preferably 1 or 2, heteroatoms each selected from O, N
and S. Each ring may have from 4 to 7, preferably 5 or 6, ring
atoms. A bicyclic heteroaromatic ring system may include a
carbocyclic ring. Examples of heteroaryl groups include pyrrole,
quinoline, isoquinoline, pyridine, pyrimidine, oxazole, thiazole,
thiadiazole, triazole, imidazole and benzimidazole.
When used herein (except for in respect of R.sup.6 and R.sup.7),
the term "mono-cyclic" means an aromatic or heteroaromatic group
having a 3 to 8 membered ring system for example phenyl, pyridine
or pyran. When used herein the term "bicyclic ring" means an
aromatic or heteroaromatic fused ring system in which at least one
of the rings is aromatic or heteroaromatic for example naphthyl,
indole, benzofuran, indene, fused phenylcyclohexane, or fused
phenyl cyclopentane.
Aryl, heterocyclyl, heteroaryl groups and mono and bicyclic ring
systems may be optionally substituted by preferably up to three
substituents. Suitable substituents include halogen, hydroxy,
C.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxyC.sub.1-6alkyl, haloC.sub.1-6alkyl,
arylC.sub.1-6alkoxy, hydroxy, nitro, cyano, azido, amino, mono- and
di-N-C.sub.1-6alkylamino, acylamino, arylcarbonylamino, acyloxy,
carboxy, carboxy salts, carboxy esters, carbamoyl, mono- and
di-N-C.sub.1-6alkylcarbamoyl, C.sub.1-6alkoxycarbonyl,
aryloxycarbonyl, ureido, guanidino, C.sub.1-6alkylguanidino,
amidino, C.sub.1-6alkylamidino, sulphonylamino, aminosulphonyl,
C.sub.1-6alkylthio, C.sub.1-6alkylsulphinyl,
C.sub.1-6alkylsulphonyl, heterocyclyl, heteroaryl, heterocyclyl
C.sub.1-6alkyl, hydroxyimino-C.sub.1-6alkyl and heteroaryl
C.sub.1-6alkyl, and combinations thereof.
Preferably the optional substituent contains a water-solubilising
group; suitable solubilising moieties will be apparent to those
skilled in the art and include hydroxy and amine groups. Even more
preferably the optional substituent includes amino, mono- or
di-C.sub.1-6alkylamino, amine containing heterocycle or hydroxy or
any combination thereof. Other preferred substituents are
C.sub.1-6alkyl and C.sub.1-6alkoxyC.sub.1-6alkyl.
When used herein the term halo represents fluoro, chloro, bromo or
iodo.
X is preferably NH or X--R.sup.1 is preferably hydrogen.
When X is NH, R.sup.1 is preferably hydrogen or C.sub.1-6alkyl.
When Y.sub.1 and Y.sub.2 are CH, X--R.sup.1 is preferably
hydrogen.
When Y.sub.2 is N, R.sup.1 is preferably hydrogen or
C.sub.1-6alkyl.
Preferably R.sup.11 is hydrogen.
Most preferably X--R.sup.1 is hydrogen
Preferably X.sup.1 or X.sub.2 is S or O, most preferably O.
A is preferably a fused 5 membered ring optionally containing up to
two heteroatoms selected from O, S and NR.sup.5, wherein R.sup.5 is
hydrogen or C.sub.1-6alkyl, which ring is optionally substituted by
up to 2 substituents selected from halogen, C.sub.1-6alkyl,
hydroxy, C.sub.1-6alkoxy or keto.
Even more preferably A is a fused 5 membered ring.
Preferably R.sup.6 and R.sup.7 are independently selected from
hydrogen, C.sub.1-6alkyl, aryl, heterocyclic and
heterocyclicC.sub.1-6alkyl wherein any of the groups except
hydrogen may be optionally substituted or R.sup.6 and R.sup.7
together with the nitrogen atom to which they are attached form a
3- to 7-membered monocyclic optionally including upto three
heteroatoms selected from O, N or S wherein said ring may be
optionally substituted.
Most preferably the compounds of the invention are of formula
(II);
##STR00003##
wherein R.sup.1, X, Y.sub.1, Y.sub.2, R.sup.3, X.sub.1, X.sub.2,
R.sup.2 and R.sup.4 are as defined for compounds of formula (I) or
pharmaceutically acceptable salts thereof.
The compounds of formula (I) preferably have a molecular weight of
less than 800.
Preferred substituents for the group Ar include halo, hydroxy,
hydroxyC.sub.1-6alkyl, hydroxyimino-C.sub.1-6alkyl and
C.sub.1-6alkoxy.
It will be understood that the invention includes pharmaceutically
acceptable derivatives of compounds of formula (I) and that these
are included within the scope of the invention.
Particular compounds according to the invention include those
mentioned in the examples and their pharmaceutically acceptable
salts. As used herein "pharmaceutically acceptable derivatives"
includes any pharmaceutically acceptable salt, ester or salt of
such ester of a compound of formula (I) which, upon administration
to the recipient, is capable of providing (directly or indirectly)
a compound of formula (I) or an active metabolite or residue
thereof.
It will be appreciated that for use in medicine the salts of the
compounds of formula (I) should be pharmaceutically acceptable.
Suitable pharmaceutically acceptable salts will be apparent to
those skilled in the art and include those described in J. Pharm.
Sci., 1977, 66, 1-19, such as acid addition salts formed with
inorganic acids e.g. hydrochloric, hydrobromic, sulfuric, nitric or
phosphoric acid; and organic acids e.g. succinic, maleic, acetic,
fumaric, citric, tartaric, benzoic, p-toluenesulfonic,
methanesulfonic or naphthalenesulfonic acid. Other salts e.g.
oxalates, may be used, for example in the isolation of compounds of
formula (I) and are included within the scope of this
invention.
The compounds of this invention may be in crystalline or
non-crystalline form, and, if crystalline, may optionally be
hydrated or solvated. This invention includes within its scope
stoichiometric hydrates as well as compounds containing variable
amounts of water.
The invention extends to all isomeric forms including stereoisomers
and geometric isomers of the compounds of formula (I) including
enantiomers and mixtures thereof e.g. racemates. The different
isomeric forms may be separated or resolved one from the other by
conventional methods, or any given isomer may be obtained by
conventional synthetic methods or by stereospecific or asymmetric
syntheses.
Since the compounds of formula (I) are intended for use in
pharmaceutical compositions it will readily be understood that they
are each preferably provided in substantially pure form, for
example at least 60% pure, more suitably at least 75% pure and
preferably at least 85%, especially at least 98% pure (% are on a
weight for weight basis). Impure preparations of the compounds may
be used for preparing the more pure forms used in the
pharmaceutical compositions.
Compounds of formula (I) are furan, pyrrole and thiophene
derivatives which may be readily prepared, using procedures
well-known to those skilled in the art, from starting materials
which are either commercially available or can be prepared from
such by analogy with well-known processes. For instance see, W.
Friedrichsen (p351, furans), R. J. Sundberg (p119, pyrroles) and J.
Nakayama (p607, thiophenes) in Comprehensive Heterocyclic Chemistry
II, volume 2, series eds. A. R. Katritzky, C. W. Rees and E. F. V.
Scriven.
Typically, compounds of this invention may be prepared by a
sequential transition metal catalysed cross-coupling procedure on a
2,3-dihalo heterocycle, as shown in Scheme 1; this is particularly
applicable for furan or thiophene derivatives, i.e. when either
X.sub.1 or X.sub.2 are O or S. For example, Suzuki coupling of
pyridine-4-boronic acid with 2,3-dibromofuran-5-carboxylic acid
t-butyl ester (1) preferentially results in the formation of the
2-(4-pyridyl) derivative (2). Subsequent Suzuki reaction with an
indanone boronic acid derivative (3, wherein PG is O, N--OMe or
another ketone protecting group) then generates the derivative (4).
Thereafter, the ester group may be converted into an amide group
using appropriate conventional functional group interconversion
procedures and the group PG converted into an hydroxyimino group as
in (5). It will also be appreciated, to one skilled in the art,
that the above cross-coupling reactions may be carried out in
reverse order giving access to the regioisomeric heterocycles
(6).
##STR00004##
wherein X.sub.1, X.sub.2, R.sup.6 and R.sup.7 are as defined for
compounds of formula (I), and PG is a protecting group.
During the synthesis of the compounds of formula (I) labile
functional groups in the intermediate compounds, e.g. hydroxy,
carboxy and amino groups, may be protected. A comprehensive
discussion of the ways in which various labile functional groups
may be protected and methods for cleaving the resulting protected
derivatives is given in for example Protective Groups in Organic
Chemistry, T. W. Greene and P. G. M. Wuts, (Wiley-Interscience, New
York, 2nd edition, 1991).
The compounds of formula (I) may be prepared singly or as compound
libraries comprising at least 2, for example 5 to 1,000 compounds,
and more preferably 10 to 100 compounds of formula (I). Libraries
of compounds of formula (I) may be prepared by a combinatorial
`split and mix` approach or by multiple parallel synthesis using
either solution phase or solid phase chemistry, by procedures known
to those skilled in the art.
Thus according to a further aspect of the invention there is
provided a compound library comprising at least 2 compounds of
formula (I), or pharmaceutically acceptable salts thereof.
Pharmaceutically acceptable salts may be prepared conventionally by
reaction with the appropriate acid or acid derivative.
The novel carboxylic esters and the corresponding acids of formula
(III) which are used as intermediates in the synthesis of the
compounds of formula (I) and (II) also form part of the present
invention:
##STR00005## wherein X, Y.sub.1, Y.sub.2, R.sup.1, R.sup.3, Ar,
X.sub.1 and X.sub.2 are as defined for compounds of formula (I) and
R is hydrogen, C.sub.1-6alkyl or arylC.sub.1-6alkyl.
As indicated above the compounds of formula (I) and their
pharmaceutically acceptable derivatives are useful for the
treatment and/or prophylaxis of disorders in which Raf kinases, in
particular B-Raf kinase, are implicated.
According to a further aspect of the invention there is provided
the use of a compound of formula (I) or a pharmaceutically
acceptable salt thereof as an inhibitor of B-Raf kinase.
As indicated above the compounds of formula (I) and their
pharmaceutically acceptable derivatives are useful in the treatment
and/or prophylaxis of disorders associated with neuronal
degeneration resulting from ischemic events, cancer, as well as
chronic neurodegeneration, pain, migraine and cardiac
hypertropy.
According to a further aspect of the invention there is provided a
method of treatment or prophylaxis of a neurotraumatic disease, in
a mammal in need thereof, which comprises administering to said
mammal an effective amount of a compound of formula (I) or a
pharmaceutically acceptable derivative thereof.
According to a further aspect of the invention there is provided
the use of a compound of formula (I) or a pharmaceutically
acceptable derivative thereof in the manufacture of a medicament
for the prophylactic or therapeutic treatment of any disease state
in a human, or other mammal, which is exacerbated or caused by a
neurotraumatic event.
Neurotraumatic diseases/events as defined herein include both open
or penetrating head trauma, such as caused by surgery, or a closed
head trauma injury, such as caused by an injury to the head region.
Also included within this definition is ischemic stroke,
particularly to the brain area, transient ischemic attacks
following coronary by-pass and cognitive decline following other
transient ischemic conditions.
Ischemic stroke may be defined as a focal neurologic disorder that
results from insufficient blood supply to a particular brain area,
usually as a consequence of an embolus, thrombi, or local
atheromatous closure of the blood vessel. Roles for stress stimuli
(such as anoxia), redox injury, excessive neuronal excitatory
stimulation and inflammatory cytokines in this area has been
emerging and the present invention provides a means for the
potential treatment of these injuries. Relatively little treatment,
for an acute injury such as these has been available.
The compounds of the invention may also be used in the treatment or
prophylaxis of cancers. It is suggested that the compounds are
effective in tumors that have activating B-Raf mutations (V599E) as
well as tumors that are activated by Ras mutation. Mutations may
occur in the Ras family members such as Kras2 with mutation G13D.
Furthermore compounds of the invention may be used in the treament
or prophylaxis of colorectal cancer and melanoma
According to a further aspect of the invention there is provided a
method of treatment or prophylaxis of a mammal who is suffering
from or susceptible to cancer, which comprises administering to
said mammal an effective amount of a compound of formula (I) or a
pharmaceutically acceptable derivative thereof.
According to a further aspect of the invention there is provided
the use of a compound of formula (I) or a pharmaceutically
acceptable derivative thereof in the manufacture of a medicament
for the prophylactic or therapeutic treatment of cancers.
The compounds of formula (I) and pharmaceutically acceptable
derivatives thereof, may be employed alone or in combination with
other therapeutic agents for the treatment of the above-mentioned
conditions. In particular, in anti-cancer therapy, combination with
other chemotherapeutic, hormonal or antibody agents is envisaged as
well as combination with surgical therapy and radiotherapy.
Combination therapies according to the present invention thus
comprise the administration of at least one compound of formula (I)
or a pharmaceutically acceptable derivative thereof, and the use of
at least one other cancer treatment method. Preferably, combination
therapies according to the present invention comprise the
administration of at least one compound of formula (I) or a
pharmaceutically acceptable derivative thereof, and at least one
other pharmaceutically active chemotherapeutic agent. These include
existing and prospective chemotherapeutic agents. The compound(s)
of formula (I) and the other pharmaceutically active
chemotherapeutic agent(s) may be administered together in a unitary
pharmaceutical composition or separately and, when administered
separately this may occur simultaneously or sequentially in any
order. Such sequential administration may be close in time or
remote in time. The amounts of the compound(s) of formula (I) and
the other pharmaceutically active chemotherapeutic agent(s) and the
relative timings of administration will be selected in order to
achieve the desired combined therapeutic effect.
Pharmaceutically active chemotherapeutic agents which can be useful
in combination with a compound of formula (I) or a pharmaceutically
acceptable derivative thereof, include but are not restricted to
the following:
(1) cell cycle specific anti-neoplastic agents include, but are not
limited to, diterpenoids such as paclitaxel and its analog
docetaxel; tubulin poisons such as taxol/taxane or vinca alkaloids
such as vinblastine, vincristine, vindesine, and vinorelbine;
epipodophyllotoxins such as etoposide and teniposide;
fluoropyrimidines such as 5-fluorouracil and fluorodeoxyuridine;
antimetabolites such as allopurinol, fludarabine, methotrexate,
cladrabine, cytarabine, mercaptopurine, gemcitabine, and
thioguanine; and camptothecins such as 9-amino camptothecin,
irinotecan, topotecan, and the various optical forms of
7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptoth-
ecin;
(2) cytotoxic chemotherapeutic agents including, but not limited
to, alkylating agents such as melphalan, chlorambucil,
cyclophosphamide, mechlorethamine, hexamethylmelamine, busulfan,
carmustine, lomustine, dacarbazine and nitrosoureas; anti-tumour
antibiotics such as doxorubicin, daunomycin, epirubicin,
idarubicin, mitomycin-C, dacttinomycin, bleomycin and mithramycin;
and platinum coordination complexes such as cisplatin, carboplatin,
and oxaliplatin; and
(3) other chemotherapeutic agents including, but not limited to,
anti-estrogens such as tamoxifen, toremifene, raloxifene,
droloxifene and iodoxyfene; progestrogens such as megestrol
acetate; aromatase inhibitors such as anastrozole, letrazole,
vorazole, and exemestane; antiandrogens such as flutamide,
nilutamide, bicalutamide, and cyproterone acetate; LHRH agonists
and antagagonists such as goserelin acetate and luprolide,
testosterone 5.alpha.-dihydroreductase inhibitors such as
finasteride; metalloproteinase inhibitors such as marimastat;
antiprogestrogens; mitoxantrone, 1-asparaginase, urokinase
plasminogen activator receptor function inhibitors; inhibitors or
c-kit and bcr/ab1 tyrosine kinases, (such as Gleevec),
immunotherapy, immunoconjugates, cytokines (such as IL-2, IFN alpha
and beta), tumor vaccines (including dendritic cell vaccines),
thalidomide, COX-2 inhibitors, glucocorticoids (such as prednisone
and decadron), radiation sensitizers, (such as temazolamide),
growth factor function inhibitors such as inhibitors of the
functions of hepatocyte growth factor; erb-B2, erb-B4, epidermal
growth factor receptor (EGFR) and platelet derived growth factor
receptors (PDGFR); inhibitors of angiogenesis such as inhibitors of
the function of Ephrin receptors (such as, EphB4), vascular
endothelial growth factor receptors (VEGFR) and the angiopoietin
receptors (Tie1 and Tie2); and other kinase inhibitors such as
inhibitors of CDK2 and CDK4.
Anti-neoplastic agents may induce anti-neoplastic effects in a
cell-cycle specific manner, i.e., are phase specific and act at a
specific phase of the cell cycle, or bind DNA and act in a non
cell-cycle specific manner, i.e., are non-cell cycle specific and
operate by other mechanisms.
According to a further aspect of the invention there is provided a
method of treatment or prophylaxis of chronic neurodegeneration,
pain, migraine or cardiac hypertrophy, in a mammal in need thereof,
which comprises administering to said mammal an effective amount of
a compound of formula (I) or a pharmaceutically acceptable
derivative thereof.
According to a further aspect of the invention there is provided
the use of a compound of formula (I) or a pharmaceutically
acceptable derivative thereof in the manufacture of a medicament
for the prophylactic or therapeutic treatment of chronic
neurodegeneration, pain, migraine or cardiac hypertrophy.
In order to use the compounds of formula (I) in therapy, they will
normally be formulated into a pharmaceutical composition in
accordance with standard pharmaceutical practice.
According to a further aspect of the invention there is provided a
pharmaceutical composition comprising a compound of formula (I) or
a pharmaceutically acceptable derivative thereof and a
pharmaceutically acceptable carrier.
The compounds of formula (I) may conveniently be administered by
any of the routes conventionally used for drug administration, for
instance, parenterally, orally, topically or by inhalation. The
compounds of formula (I) may be administered in conventional dosage
forms prepared by combining it with standard pharmaceutical
carriers according to conventional procedures. The compounds of
formula (I) may also be administered in conventional dosages in
combination with a known, second therapeutically active compound.
These procedures may involve mixing, granulating and compressing or
dissolving the ingredients as appropriate to the desired
preparation. It will be appreciated that the form and character of
the pharmaceutically acceptable carrier is dictated by the amount
of compound of formula (I) with which it is to be combined, the
route of administration and other well-known variables. The
carrier(s) must be "acceptable" in the sense of being compatible
with the other ingredients of the formulation and not deleterious
to the recipient thereof.
The pharmaceutical carrier employed may be, for example, either a
solid or liquid. Exemplary of solid carriers are lactose, terra
alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium
stearate, stearic acid and the like. Exemplary of liquid carriers
are syrup, peanut oil, olive oil, water and the like. Similarly,
the carrier or diluent may include time delay material well known
to the art, such as glyceryl mono-stearate or glyceryl distearate
alone or with a wax.
A wide variety of pharmaceutical forms can be employed. Thus, if a
solid carrier is used, the preparation can be tableted, placed in a
hard gelatin capsule in powder or pellet form or in the form of a
troche or lozenge. The amount of solid carrier will vary widely but
preferably will be from about 25 mg to about 1 g. When a liquid
carrier is used, the preparation will be in the form of a syrup,
emulsion, soft gelatin capsule, sterile injectable liquid such as
an ampoule or nonaqueous liquid suspension.
The compounds of formula (I) are preferably administered
parenterally, that is by intravenous, intramuscular, subcutaneous,
sublingual, intranasal, intrarectal, intravaginal or
intraperitoneal administration. The intravenous form of parenteral
administration is generally preferred. The compounds may be
administered as a bolus or continuous infusion e.g. of 6 hours up
to 3 days. Appropriate dosage forms for such administration may be
prepared by conventional techniques.
The compounds of formula (I) may also be administered orally.
Appropriate dosage forms for such administration may be prepared by
conventional techniques.
The compounds of formula (I) may also be administered by
inhalation, that is by intranasal and oral inhalation
administration. Appropriate dosage forms for such administration,
such as aerosol formulations, may be prepared by conventional
techniques.
The compounds of formula (I) may also be administered topically,
that is by non-systemic administration. This includes the
application of the inhibitors externally to the epidermis or the
buccal cavity and the instillation of such a compound into the ear,
eye and nose, such that the compound does not significantly enter
the blood stream.
For all methods of use disclosed herein the daily oral dosage
regimen will preferably be from about 0.1 to about 80 mg/kg of
total body weight, preferably from about 0.2 to 30 mg/kg, more
preferably from about 0.5 to 15 mg/kg. The daily parenteral dosage
regimen about 0.1 to about 80 mg/kg of total body weight,
preferably from about 0.2 to about 30 mg/kg, and more preferably
from about 0.5 to 15 mg/kg. The daily topical dosage regimen will
preferably be from 0.1 mg to 150 mg, administered one to four,
preferably two or three times daily. The daily inhalation dosage
regimen will preferably be from about 0.01 mg/kg to about 1 mg/kg
per day. It will also be recognized by one of skill in the art that
the optimal quantity and spacing of individual dosages of the
inhibitors will be determined by the nature and extent of the
condition being treated, the form, route and site of
administration, and the particular patient being treated, and that
such optimums can be determined by conventional techniques. It will
also be appreciated by one of skill in the art that the optimal
course of treatment, i.e., the number of doses of the inhibitors
given per day for a defined number of days, can be ascertained by
those skilled in the art using conventional course of treatment
determination tests. In the case of pharmaceutically acceptable
salts the above figures are calculated as the parent compound of
formula (I).
No toxicological effects are indicated/expected when a compound of
formula (I) is administered in the above mentioned dosage
range.
All publications, including but not limited to patents and patent
applications, cited in this specification are herein incorporated
by reference as if each individual publication were specifically
and individually indicated to be incorporated by reference herein
as though fully set forth.
The following Examples illustrate the preparation of
pharmacologically active compounds of the invention and the
following Descriptions illustrate the preparation of intermediates
used in the preparation of these compounds.
Abbreviations used herein are as follows;
THF means tetrahydrofuran. DMF means N,N-Dimethylformamide.
Description 1: 1-Methoxyimino-indan-5-boronic acid
Step 1. 5-Bromoindan-1-one O-methyl oxime
To a solution of 5-bromoindan-1-one (100 g, 0.47 mol) in ethanol
(650 ml) under argon was added methoxylamine hydrochloride (198 g,
2.38 mol) and pyridine (125 ml). The mixture was heated under
reflux for 2.5 hours, cooled to room temperature and poured into
saturated aqueous sodium hydrogen carbonate solution. The mixture
was extracted with ethyl acetate and the organic phase dried and
concentrated in vacuo. The crude material was recrystallised from
isopropanol to afford the title compound (110 g, 97%); .sup.1H NMR
(CDCl.sub.3) 7.52 (1H, d, J 8.3 Hz), 7.43 (1H, d, J 1 Hz), 7.35
(1H, dd, J 8.3, 1 Hz), 3.97 (3H, s), 2.99 (2H, m), 2.85 (2H,
m).
Step 2. 1-Methoxyimino-indan-5-boronic acid
A solution of the product of Example 1 Step 1 (48.0 g, 0.2 mol) in
tetrahydrofuran (1 L) at -78.degree. C. under argon atmosphere was
treated dropwise with n-butyl lithium (138 ml, 1.6M in hexanes,
0.22 mol). After stirring at -78.degree. C. for 30 minutes
trimethyl borate (49 ml, 0.44 mol) was added and the solution
warmed to room temperature and stirred for 16 hours. The mixture
was concentrated in vacuo, acidified to pH1 with 5N hydrochloric
acid and stirred at room temperature for 1 hour. The mixture was
then basified with 40% sodium hydroxide and the solution washed
three times with diethyl ether. The aqueous phase was re-acidified
to pH1 and the mixture was extracted five times with ethyl acetate.
The organic extracts were combined washed with brine, dried and
evaporated in vacuo. The residue was triturated with hexane,
filtered, washed with hexane and then a small amount of ether to
afford the title compound (23.6 g, 58%); MS(AP-) m/e 204
[M-H]-.
Description 2:
4-(1-Oxo-indan-5-yl)-5-pyridin4-yl-furan-2-carboxylic acid
hydrochloride
Step 1. 4-Bromo-5-pyridin-4-yl-furan-2-carboxylic acid tert-butyl
ester
tert-Butyl 4,5-dibromo-2-furancarboxylate (H. Muratake et al, Chem.
Pharm. Bull., 1997, 45, 799) (9.78 g, 30 mmol), 4-pyridyl boronic
acid (M. Lamothe et al, J. Med. Chem., 1997, 40, 3542) (4.06 g, 33
mmol), potassium carbonate (24.8 g, 180 mmol), triphenylphosphine
(786 mg, 3 mmol) and palladium (II) acetate (337 mg, 1.5 mmol) were
dissolved in ethylene glycol dimethyl ether (150 ml) and water (75
ml). The mixture was heated at reflux for 18 hours with vigorous
stiring, cooled then filtered through celite pad, which was
thoroughly washed with ethyl acetate. The filtrate was then washed
with saturated sodium bicarbonate solution, water (.times.3) and
brine, then dried over magnesium sulfate. The solution was
evaporated in vacuo and the residue purified by silica gel
chromatography to afford the title product (5.32 g, 55%); MS (AP-)
m/e 323/325 [M-H].sup.-.
Step 2.
4-(1-Methoxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid tert-butyl ester
The product of Step 1 (5.32 g, 16.4 mmol), the product of
Description 1 (4.03 g, 19.7 mmol), potassium carbonate (13.6 g,
98.4 mmol), triphenylphosphine (365 mg, 1.64 mmol) and palladium
acetate (184 mg, 0.8 mmol) were dissolved in ethylene glycol
dimethyl ether (100 ml) and water (50 ml). The mixture was then
heated under reflux for 5 hours, cooled and filtered through a pad
of celite. The filtrate was then washed with saturated sodium
bicarbonate solution, water and brine. The solution was dried over
magnesium sulfate and evaporated in vacuo and the residue purified
by silica gel chromatography eluting with a mixture of ethyl
acetate/hexane (1:1) to afford the title product (4.74 g, 71%); MS
(AP-) m/e 403 [M-H].sup.-.
Step 3.
4-(1-Methoxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid trifluoroacetic acid salt
The product from Step 2 (4.74 g, 11.7 mmol) was dissolved in
trifluoroacetic acid (50 ml) and dichloromethane (50 ml). The
solution was stirred at room temperature for 3 hours and then
evaporated in vacuo, azeotroping three times with dichloromethane.
The resulting solid was triturated with diethyl ether, filtered and
dried to afford the title product (5.46 g), which was used without
further purification; MS (AP-) m/e 347 [M-H].sup.-.
Step 4. 4-(1-Oxo-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic acid
hydrochloride salt
The product from Step 3 (5.46 g, 11.7 mmol) was suspended in 5M
hydrochloric acid (50 ml), dioxane (50 ml) and acetone (10 ml). The
suspension was heated to reflux for 30 minutes, whereupon the
suspended solids dissolved. The heating was continued for a further
90 minutes before the mixture was cooled, diluted with acetone (100
ml) and evaporated in vacuo to a wet solid. The solid was dried by
repeatedly (.times.3) suspending in toluene and evaporating to
dryness.
The residue was then triturated with diethyl ether, filtered and
dried in vacuo to afford the title compound (4.10 g, 97%); MS (ES+)
m/e 320 [M+H].sup.+.
Description 3: 5-(Oxo-indan-5-yl)-4pyridin-4-yl-furan-2-carboxylic
acid hydrochloride
Step 1. 4Bromo-5-(1-methoxyimino-indan-5-yl)-furan-2-carboxylic
acid tert-butyl ester
The title compound (5.83 g, 46%) was prepared from tert-butyl
4,5-dibromo-2-furancarboxylate (H. Muratake et al, Chem. Pharm.
Bull., 1997, 45, 799), (10.104 g, 30.99 mmol) and the product from
Description 1 Step 2 (6.355 g, 30.99 mmol) by the general procedure
of Description 2 Step 1; MS (AP+ve) m/e 406/408 [M+H].sup.+.
Step 2.
5-(1-Methoxyimino-indan-5-yl-4-pyridin-4-yl-furan-2-carboxylic acid
tert-butyl ester
4-Tributylstannyl pyridine (13.659 g, 37.11 mmol) and dichlorobis
(triphenylphosphine)palladium II (3.472 g, 4.948 mmol) were added
to a solution of the product from Step1 (10.05 g, 24.737 mmol), in
toluene (100 ml) and the mixture heated under reflux for 4 days.
The reaction was then cooled and filtered through a pad of celite,
and washed thoroughly with ethyl acetate. The organics were washed
with saturated sodium bicarbonate solution, water (.times.3) and
brine, dried over anhydrous magnesium sulfate and then evaporated
in vacuo. The crude residue was chromatographed on silica gel
eluting with a mixture of ethyl acetate/hexane (1:1) to afford the
title compound (6.82 g, 68%); MS (AP+ve) m/e 405 [M+H].sup.+.
Step 3. 5-(1-Oxo-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic acid
hydrochloride salt
The title compound (2.32 g, 84%) was prepared from the product from
step 2 (3.17 g, 7.83 mmol) by the general 2 step method of
description 2, steps 3 and 4; MS (ES.sup.+) m/e 320
[M+H].sup.+.
EXAMPLE 1
5-[5-(1-Morpholin-4-yl-methanoyl)-2-pyridin-4-yl-furan-3-yl]-indan-1-one
oxime
##STR00006## Step 1.
5-{5-(1-Morpholin-4-yl-methanoyl)-2-pyridin-4-yl-furan-3-yl]-indan-1-one
The product of Description 2 (178 mg, 0.5 mmol),
N-cyclohexylcarbodiimide,N'-methyl polystyrene resin (1.8 mmol/g)
(555 mg, 1 mmol), and 1-hydroxybenzotriazole hydrate (135 mg, 1
mmol) were suspended in DMF (5 ml) and then treated with
triethylamine (0.083 ml, 0.6 mmol) and morpholine (0.053 ml, 0.6
mmol). The reaction was stirred at room temperature for 16 hours
and applied to a 10 g SCX cartridge (Varian Mega Bond Elute). The
cartridge was washed with methanol and then a mixture of 0.880
ammonia/methanol (1:9) solution. The product containing fractions
were combined, evaporated in vacuo and the residue purified by
silica gel chromatography eluting with a mixture of 0.880
ammonia/ethanol/dichloromethane (1:9:90) to afford the title
compound (137 mg, 71%); MS (ES+) m/e 389 (M+H).sup.+.
Step 2.
5-[5-(1-Morpholin-4-yl-methanoyl)-2-pyridin-4-yl-furan-3-yl]-inda-
n-1-one oxime
The product from Step 1 (137 mg, 0.35 mmol) was dissolved in
ethanol (10 ml) and treated with hydroxylamine (1 ml, 50% aqueous
solution) and the solution heated to reflux for 1 hour. After
cooling, the reaction mixture was concentrated and the residue
tritrurated with diethyl ether, filtered and the solid dried in
vacuo. Purification of the solid by silica gel chromatography
eluting with a mixture of 0.880 ammonia/ethanol/dichloromethane
(1:9:90) afforded the title compound (43 mg, 30%); MS (ES+) m/e 404
(M+H).sup.+.
The following examples were prepared by the general two-step method
described in Example 1.
TABLE-US-00001 Example Amine Characterisation 2
5-[5-(1-Piperidin-1-yl-methanoyl)- piperidine MS(AP+) m/e
2-pyridin-4-yl-furan-3-yl]-indan- 402 [M + H].sup.+ 1-one oxime 3
4-(1-Hydroxyimino-indan-5-yl)-5- 4-(2- MS(AP+) m/e
pyridin-4-yl-furan-2-carboxylic aminoethyl)morpholine 447 [M +
H].sup.+ acid (2-morpholin-4-yl-ethyl)- amide 4
5-(5-{1-[4-(2-Methoxy-ethyl)- 1-(2-methoxy-ethyl)- MS(AP+) m/e
piperazin-1-yl]-methanoyl}-2- piperazine, (Kazunori et al., 461 [M
+ H].sup.+ pyridin-4-yl-furan-3-yl)-indan-1- Chem. Pharm. Bull.
1993, one oxime 41,148) 5 4-(1-Hydroximino-indan-5-yl)-5-
1-methyl-4- MS(ES+) m/e 445 pyridin-4-yl-furan-2-carboxylic
(methylamino)piperidine [M + H].sup.+ acid
methyl-(1-methyl-piperidin- 4-yl)-amide 6
5-[5-(1-[1,4']Bipiperidinyl-1'-yl- 4-piperidinopiperidine MS(ES+)
m/e 485 methanoyl)-2-pyridin-4-yl-furan- [M + H].sup.+
3-yl]-indan-1-one oxime 7 5-(5-{1-[4-(4-Chloro-benzyl)-
1-(4-chlorobenzyl)- MS(ES+) m/e piperazin-1-yl]-methanoyl}-2-
piperazine 527/529 [M + H].sup.+ pyridin-4-yl-furan-3-yl)-indan-1-
one oxime 8 5-{5-[1-(4-Cyclohexyl-piperazin- 1-cyclohexylpiperazine
MS(ES+) m/e 485 1-yl)-methanoyl]-2-pyridin-4-yl- [M + H].sup.+
furan-3-yl}-indan-1-one oxime 9 5-{5-[1-(4-Methyl-piperazin-1-yl)-
1-methylpiperazine MS(ES+) m/e 417 methanoyl]-2-pyridin-4-yl-furan-
[M + H].sup.+ 3-yl}-indan-1-one oxime 10
5-(5-{1-[4-(2-Oxo-2-pyrrolidin-1- 1-(2-(piperazin-1-yl)- MS(ES+)
m/e 514 yl-ethyl)-piperazin-1- acetyl)-pyrrolidine [M + H].sup.+
yl]methanoyl}-2-pyridin-4-yl- furan-3-yl)-indan-1-one oxime 11
4-(1-Hydroxyimino-indan-5-yl)-5- 4- MS(ES+) m/e 467
pyridin-4-yl-furan-2-carboxylic dimethylaminobenzylamine [M +
H].sup.+ acid 4-dimethyamino-benzylamide 12
4-(1-Hydroxyimino-indan-5-yl)-5- N,N,N'- MS(ES+) m/e 419
pyridin-4-yl-furan-2-carboxylic trimethylethylenediamine [M +
H].sup.+ acid (2-dimethylaminoethyl)- methylamide 13
4-[({1-[4-(1-Hydroxyimino-indan- tert-butyl-4-(aminomethyl)-
MS(AP+) m/e 5-yl)-5-pyridin-4-yl-furan-2-yl]-
tetrahydropyridin-1-(2H)- 531 [M + H].sup.+
methanoyl}-amino)-methyl]- carboxylate piperidine-1-carboxylic acid
tert- butyl ester 14 4-(1-Hydroxyimino-indan-5-yl)-5-
N,N-dimethlyethylene MS(AP+) m/e pyridin-4-yl-furan-2-carboxylic
diamine 405 [M + H].sup.+ acid (2-dimethylamino-ethyl)- amide 15
5-(2-Pyridin-4-yl-5-{1-[4-(2- 1-(2-(1-pyrrolidyl)-ethyl)- MS(AP+)
m/e pyrrolidin-1-yl-ethyl)-piperazin-1- piperazine 500 [M +
H].sup.+ yl]-methanoyl}-furan-3-yl)-indan- 1-one oxime 16
4-(1-Hydroxyimino-indan-5-yl)-5- (1-piperidin-1-yl- MS(AP+) m/e
pyridin-4-yl-furan-2-carboxylic cyclohexyl)-methylamine 513 [M +
H].sup.+ acid (1-piperidin-4-yl- cyclohexylmethyl)-amide 17
4(1-Hydroxyimino-indan-5-yl)-5- 4-(2-dimethylamino- MS(AP+) m/e
pyridin-4-yl-furan-2-carboxylic ethoxy) phenylamine. (Paul 497 [M +
H].sup.+ acid [4-(2-dimethyl amino- et al., J. Med. Chem., 1993,
ethoxy)-phenyl]-amide 36(19), 2716) 18
4-(1-Hydroxyimino-indan-5-yl)-5- N-(2-aminoethyl)- MS(AP+) m/e
pyridin-4-yl-furan-2-carboxylic pyrrolidine 431 [M + H].sup.+ acid
(2-pyrrolidin-1-yl-ethyl)- amide 19
4-(1-Hydroxyimino-indan-5-yl)-5- 1-(3-aminopropyl)-4- MS(AP+) m/e
pyridin-4-yl-furan-2-carboxylic methylpiperazine 474 [M + H].sup.+
acid [3-(4-methyl-piperazin-1-yl)- propyl]-amide 20
5-{2-Pyridin-4-yl-5-[1-(4- 4-pyrrolidinopiperidine MS(AP+) m/e
pyrrolidin-1-yl-piperidin-1-yl)- 471 [M + H].sup.+
methanoyl}-furan-3-yl}-indan-1- one oxime 21
5-(5-{1-[4-(2-Dimethylaminoethyl)- 1-(2- MS(AP+) m/e
piperazin-1-yl]- aminomethyl)piperidine 445 [M + H].sup.+
methanoyl}-2-pyridin-4-yl-furan- 3-yl)-indan-1-one oxime 22
5-(5-{1-[4-(2-Dimethylaminoethyl)- 1-(2-dimethylaminoethyl)-
MS(AP+) m/e piperazin-1-yl]- piperazine 474 [M + H].sup.+
methanoyl}-2-pyridin-4-yl-furan- 3-yl)-indan-1-one oxime 23
4-(1-Hydroxyimino-indan-5-yl)-5- N,N-diisopropylethylene MS(AP+)
m/e pyridin-4-yl-furan-2-carboxylic diamine 461 [M + H].sup.+ acid
(2-diisopropyl amino-ethyl)- amide 24
4-(1-Hydroxyimino-indan-5-yl)-5- (1R,5S)-8-methyl-8- MS(AP+) m/e
pyridin-4-yl-furan-2-carboxylic azabicyclo[3.2.1]oct-3-yl 456 [M +
H].sup.+ acid ((1R,5S)-8-methyl-8- amine
azabicyclo[3.2.1]oct-3-yl)amide 25 4-(1-Hydroximino-indan-5-yl)-5-
((1R,5S)-9-methyl-9- MS(AP+) m/e pyridin-4-yl-furan-2-carboxylic
azabicyclo[3.3.1]non-3-yl) 471 [M + H].sup.+ acid
((1R,5S)-9-methyl-9- amine. (Bermundez et al.,
azabicyclo[3.3.1]non-3-yl)amide J. Med. Chem., 1990, 33, 7
1924)
EXAMPLE 26
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid methyl-piperidin-4-yl-amide
##STR00007## Step 1.
4-({1-[4-(1-Oxo-indan-5-yl)-5-pyridin-4-yl-furan-2-yl]-methanoyl}-amino)--
piperidine-1-carboxylic acid tert-butyl ester
The title compound (122 mg, 49%) was obtained from the product from
Description 2 (178 mg, 0.5 mmol) and
4-amino-piperidine-1-carboxylic acid tert-butyl ester (110 mg, 0.55
mmol) by the general method of example 1 step 1; MS (ES+) m/e 389
(M+H).sup.+.
Step 2.
4-(1-Hydroxyimino-indan-5-yl)-5-pyridin-4-yl-furan-2-carboxylic
acid methyl-piperidin-4-yl-amide
The product from Step 1 (122 mg, 0.24 mmol) was stirred in
trifluoroacetic acid (5 ml) and dichloromethane (5 ml) at room
temperature for 2 hours and the solution was then co-evaporated
three times with dichloromethane. The resulting residue was treated
according to the general method of Example 1 Step 2 to give the
title compound (0.02 g, 20%); MS (AP+) m/e 416 (M+H).sup.+.
The following examples were prepared by the general method
described in Example 26.
TABLE-US-00002 Example Amine Characterisation 27
5-[5-(1-Piperazin-1-yl- tert-butyl 1-piperazine MS(AP+) m/e
methanoyl)-2-pyridin-4-yl)-furan carboxylate 403 [M + H].sup.+
3-yl]-indan-1-one oxime 28 4-(1-Hydroxyimino-indan-5-yl)-5-
1-tert-butoxycarbonyl-4- MS(AP+) m/e
pyridin-4-yl-furan-2-carboxylic methylaminopiperidine 431 [M +
H].sup.+ acid methyl-piperidin-4-yl-amide PCT Int. Appl (1999) WO
9964394
EXAMPLE 29
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-dimethylamino-ethyl)-methyl-amide
##STR00008## Step 1.
5-(1-Oxo-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic acid
(2-dimethylamino-ethyl)-methyl-amide
The product from Description 3 (200 mg, 0.562 mmol),
N-cyclohexylcarbodiimide N'-methyl polystyrene resin (1.8 mmol/g)
(938 mg, 1.686 mmol), and 1-hydroxybenzotriazole hydrate (228 mg,
1.686 mmol) were suspended in dimethylformamide (5 ml) and
dichloromethane (3 ml) and treated with triethylamine (58 mg, 0.562
mmol) and N,N,N-trimethylethlenediamine (172 mg, 1.686 mmol). The
reaction was stirred at room temperature for 16 hours and then
applied to a 10 g SCX cartridge (Varian Mega Bond Elute). The
cartridge was washed with methanol and then a mixture of 0.880
ammonia/methanol (1:10). The product containing fractions were
combined, evaporated in vacuo, and the residue chromatographed on
silica gel eluting with a mixture of 0.880
ammonia/methanol/dichloromethane (1:9:90) to afford the title
compound (180 mg, 80%); MS (AP+) m/e 404 (M+H).sup.+.
Step 2.
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-dimethylamino-ethyl)-methyl-amide
The title compound (110 mg, 59%) was prepared from the product of
Step 1 using the method of Example 1 Step 2; MS (AP+ve): m/e 419
(M+H).sup.+.
The following examples were prepared by the general two-step method
described in Example 29. Varying quantities of triethylamine were
added to the reactions in Step 1 as indicated:
TABLE-US-00003 Example Amine Et.sub.3N Characterisation 30
5-(1-Hydroxyimino-indan-5- 1-methyl-4- 1 equivalent MS(AP+) m/z 445
yl)-4-pyridin-4-yl-furan-2- (methylamino) [M + H].sup.+ carboxylic
acid methyl-(1- piperidine methyl-piperidin-4-yl)-amide 31
5-[5-(1-[1,4']Bipiperidinyl-1'- 4-piperidinopiperidine 1 equivalent
MS(AP+) m/z 485 yl-methanoyl)-3-pyridin-4-yl- [M + H].sup.+
furan-2-yl]-indan-1-one oxime 32 5-(5-{1-[4-(4-Chloro-benzyl)-
1-(4-chlorobenzyl)- 0 equivalent MS(AP+) m/z
piperazin-1-yl]-methanoyl}-3- piperazine 527/529 [M + H].sup.+
pyridin-4-yl-furan-2-yl)-indan- 1-one oxime 33
5-{5-[1-(4-Cyclohexyl- 1- 0 equivalent MS(ES+) m/e 485
piperazin-1-yl)-methanoyl]-3- cyclohexylpiperazine [M + H].sup.+
pyridin-4-yl-furan-2-yl}-indan- 1-one oxime 34
5-{5-[1-(4-Methyl-piperazin-1- 1-methylpiperazine 0 equivalent
MS(ES+) m/e 417 yl)-methanoyl]-3-pyridin-4-yl- [M + H].sup.+
furan-2-yl}-indan-1-one oxime 35 5-(1-Hydroxyimino-indan-5- 1- 0
equivalent MS(ES+) m/e 431 yl)-4-pyridin-4-yl-furan-2-
aminoethylpyrrolidine [M + H].sup.+ carboxylic acid
(2-pyrrolidin-1- yl-ethyl)-amide 36 5-(5-{1-[4-(2-Oxo-2-pyrrolidin-
1-(2-(piperazin-1-yl)- 0 equivalent MS(ES+) m/e 514
1-yl-ethyl)-piperazin-1-yl]- acetyl)-pyrrolidine [M + H].sup.+
methanoyl}-3-pyridin-4-yl- furan-2-yl)-indan-1-one oxime 37
5-(1-Hydroxyimino-indan-5- 4-dimethylamino 2 equivalents MS(ES+)
m/e 467 yl)-4-pyridin-4-yl-furan-2- benzylamine [M + H].sup.+
carboxylic acid 4- dimethylamino-benzylamide
EXAMPLE 38
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-diisopropylamino-ethyl)-amide
##STR00009## Step 1.
5-(1-Oxo-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic acid
(2-diisopropylamino-ethyl)-amide
The product from Description 3 (250 mg, 0.703 mmol),
N-cyclohexylcarbodiimide N'-methyl polystyrene resin (1.7 mmol/g)
(1.240 g, 2.190 mmol) and 1-hydroxybenzotriazole hydrate (104 mg,
0.773 mmol) were suspended in tetrahydrofuran (7 ml) and
dichloromethane (3 ml) and treated with triethylamine (78 mg, 0.773
mmol) and N,N,diisopropylethylene diamine (122 mg, 0.846 mmol). The
reaction was stirred at room temperature for 16 hours, filtered and
the filtrate evaporated in vacuo and the residue chromatographed on
silica gel eluting with a mixture of 0.880
ammonia/methanol/dichloromethane (1/9/90) to afford the title
compound (187 mg, 60%); MS (ES+) m/e 446 (M+H).sup.+.
Step 2.
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-dimethylamino-ethyl)-methyl-amide
The title compound (0.051 g, 26%) was prepared from the product of
Step 1 using the method of Example 1 Step 2; MS (AP+ve): m/e 461
(M+H).sup.+.
The following examples were prepared by the general two step method
as described in example 38.
TABLE-US-00004 Example Amine Characterisation 39A
5-(1-Hydroxyimino-indan- N.sup.1,N.sup.1-Diisopropyl-ethane-
MS(ES+) m/e 5-yl)-4-pyridin-4-yl-furan- 1,2-diamine 461 [M +
H].sup.+ 2-carboxylic acid (2- diisopropylamino-ethyl)- amide 39B
4-[({1-[5-(1- 4-Aminomethyl-piperidine-1- MS(ES+) m/e
Hydroxyimino-indan-5- carboxylic acid tert-butyl ester 531 [M +
H].sup.+ yl)-4-pyridin-4-yl-furan-2- yl]-methanoyl}-amino)-
methyl]-piperidine-1- carboxylic acid tert-butyl ester 40
5-(3-Pyridin-4-yl-5-{1-[4- 1-(2-Pyrrolidin-1-yl-ethyl)- MS(ES+) m/e
(2-pyrrolidin-1-yl-ethyl)- piperazine 500 [M + H].sup.+
piperazin-1-yl]- methanoyl}-furan-2-yl)- indan-1-one oxime 41
5-(1-Hydroxyimino-indan- 2-Amino-ethanol MS(ES+) m/e
5-yl)-4-pyridin-4-yl-furan- 378 [M + H].sup.+ 2-carboxylic acid (2-
hydroxy-ethyl)-amide 42 5-(1-Hydroxyimino-indan-
2-Pyrrolidin-1-yl-ethylamine MS(ES+) m/e
5-yl)-4-pyridin-4-yl-furan- 431 [M + H].sup.+ 2-carboxylic acid (2-
pyrrolidin-1-yl-ethyl)- amide 43 5-(1-Hydroxyimino-indan-
N.sup.1,N.sup.1-Dimethyl-ethane-1,2- MS(ES+) m/e
5-yl)-4-pyridin-4-yl-furan- diamine 405 [M + H].sup.+ 2-carboxylic
acid (2- dimethylamino-ethyl)- amide 44 5-(1-Hydroxyimino-indan-
3-(4-Methyl-piperazin-1-yl)- MS(ES+) m/e 5-yl)-4-pyridin4-yl-furan-
propylamine 474 [M + H].sup.+ 2-carboxylic acid [3-(4-
methyl-piperazin-1-yl)-propyl]- amide 45 5-(1-Hydroxyimino-indan-
C-(1-Piperidin-1-yl- MS(ES+) m/e 5-yl)-4-pyridin-4-yl-furan-
cyclohexyl)-methylamine 513 [M + H].sup.+ 2-carboxylic acid (1-
piperidin-1-yl- cyclohexylmethyl)-amide 46 5-(1-Hydroxyimino-indan-
4-(2-Dimethylamino-ethoxy)- MS(ES+) m/e 5-yl)-4-pyridin-4-yl-furan-
phenylamine 497 [M + H].sup.+ 2-carboxylic acid [4-(2-
dimethylamino-ethoxy)- phenyl]-amide 47 5-(1-Hydroxyimino-indan-
N.sup.1,N.sup.1-Dimethyl-propane-1,3- MS(ES+) m/e
5-yl)-4-pyridin-4-yl-furan- diamine 419 [M + H].sup.+ 2-carboxylic
acid (3- dimethylamino-propyl)- amide 48 5-(1-Hydroxyimino-indan-
2-Piperidin-1-yl-ethylamine MS(ES+) m/e 5-yl)-4-pyridin-4-yl-furan-
445 [M + H].sup.+ 2-carboxylic acid (2-
piperidin-1-yl-ethyl-amide
EXAMPLE 49
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-amino-ethyl)-amide
##STR00010## Step 1:
5-(1-Oxo-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic acid
(2-amino-ethyl)-amide
The title compound (0.107 g, 33%) was prepared from the product of
Description 3 (0.25 g, 0.703 mmol) and (2-amino-ethyl)-carbamic
acid tert-butyl ester (135 mg, 0.843 mmol) using the method of
Example 38 Step 1; MS (AP+) m/e 462 (M+H).sup.+.
Step 2:
5-(1-Hydroxyimino-indan-5-yl)-4-pyridin-4-yl-furan-2-carboxylic
acid (2-amino-ethyl)-amide
The title compound (0.037 g, 23%) was prepared from the product of
Step 1 using the method of Example 26 Step 2; MS (ES-ve): m/e 375
[M-H].sup.-.
The following compounds were prepared by the general two step
method as described in Example 49.
TABLE-US-00005 Example Amine Characterisation 50
5-(1-Hydroxyimino-indan- 4-Amino-piperidine-1- MS(ES+) m/e
5-yl)-4-pyridin-4-yl-furan- carboxylic acid tert-butyl ester 417 [M
+ H].sup.+ 2-carboxylic acid piperidin-4- ylamide 51 (+/-)
5-(1-Hydroxyimino-indan- (+/-) 3-Amino-pyrrolidine-1- MS(ES+) m/e
5-yl)-4-pyridin-4-yl- carboxylic acid tert-butyl ester 403 [M +
H].sup.+ furan-2-carboxylic acid pyrrolidin-3-ylamide
EXAMPLE 52
5-(1-Hydroximino-indan-5-yl)-4-pyridin-4-yl-thiophene-2-carboxylic
acid methyl-(1-methyl-piperidin-4-yl)-amide
##STR00011## Step 1. 4,5-Dibromo-thiophene-2-carboxylic acid
methyl-(1-methyl-piperidin-4-yl)-amide
4,5-Dibromothiophene-2-carboxylic acid (2.86 g, 10 mmol) was
dissolved in dichloromethane (50 ml) and treated dropwise at
0.degree. C. with oxalyl chloride (2.61 ml, 30 mmol).
N,N-Dimethylfomamide (5 drops) was added and the solution stirred
at room temperature for 3 hours. The solution was evaporated in
vacuo and co-evaporated with dichloromethane (.times.3) to afford
crude acid chloride, 3.04 g. The solid was dissolved in THF (10 ml)
and treated with a solution of 1-methyl-4-(methylamino)piperidine
(1.28 g, 10 mmol) and triethylamine (1.21 g, 12 mmol) in THF (20
ml). The mixture was stirred at room temperature for 16 hours,
evaporated in vacuo and the residue partitioned between ethyl
acetate and saturated sodium bicarbonate solution. The organic
layer was washed three times with water, then brine, dried over
magnesium sulfate and evaporated in vacuo to afford the title
product (3.41 g, 90%) which was used without further purification;
MS (ES+) m/e 397/399/401 [M+H].sup.+
Step 2.
4-Bromo-5-(1-methoxyimino-indan-5-yl)-thiophene-2-carboxylic acid
methyl-(1-methyl-piperidin-4-yl)-amide
The product of Step 1 (3.41 g, 9 mmol), the product of Description
1 (1.85 g, 9 mmol), potassium carbonate (7.46 g, 54 mmol),
triphenylphosphine (240 mg, 0.9 mmol), and palladium acetate (100
mg, 0.45 mmol) were dissolved in ethylene glycol dimethyl ether (50
ml) and water (25 ml). The biphasic solution was heated at reflux
for 36 hours, with vigorous stirring, cooled, then filtered through
a pad of celite, which was thoroughly washed with ethyl acetate.
The two phase filtrate was separated and the organic phase washed
with saturated sodium bicarbonate solution, water (.times.3) and
brine. The solution was dried over magnesium sulfate and evaporated
in vacuo to a crude solid (4.9 g), which was purified by silica gel
chromatography (1:9:90 0.880 ammonia:ethanol:dichloromethane), to
afford the title product, (670 mg, 16%); MS (AP+) m/e 477/479
[M+H].sup.+
Step 3.
5-(1-methoxyimino-indan-5-yl)-4-pyridin-4-yl-thiophene-2-carboxyl-
ic acid methyl-(1-methyl-piperidin-4-yl)-amide
The product of Step 2 (670 mg, 1.4 mmol), 4-pyridyl boronic acid
(M. Lamothe et al J. Med. Chem., 1997, 40, 3542) (189 mg, 1.5
mmol), potassium carbonate (1.17 g, 8.5 mmol), triphenylphosphine
(37 mg, 0.14 mmol), and palladium acetate (16 mg, 0.07 mmol) were
dissolved in ethylene glycol dimethyl ether (15 ml) and water (5
ml). The biphasic solution was heated at reflux for 16 hours, with
vigorous stirring, cooled, then filtered through a pad of celite,
which was thoroughly washed with ethyl acetate. The two phase
filtrate was separated and the organic phase washed with saturated
sodium bicarbonate solution, water (.times.3) and brine. The
solution was dried over magnesium sulfate and evaporated in vacuo
to a crude solid (691 mg), which was purified by silica gel
chromatography (1:9:90 0.880 ammonia:ethanol:dichloromethane), to
afford the title product, (476 mg, 72%); MS (AP+) m/e 475
[M+H].sup.+
Step 4. 5-(1-Oxo-indan-5-yl)-4-pyridin-4-yl-thiophene-2-carboxylic
acid methyl-(1-methyl-piperidin-4-yl)-amide
The product of step 3 (476 mg 1 mmol) was dissolved in 5N
hydrochloric acid (10 ml), dioxane (10 ml) and acetone (5 ml) and
heated at 80.degree. C. for 2 hours. The resulting solution was
concentrated in vacuo and co-evaporated with ethanol (.times.3) to
afford a crude solid. This was partitioned between saturated sodium
bicarbonate solution and ethyl acetate. The aqueous phase was
re-extracted with ethyl acetate (.times.2), and the combined
organic extracts were washed with water (.times.3), brine, dried
and evaporated in vacuo to a crude solid. This was purified by
column chromatography (1:9:40 0.880 ammonia:thanol:dichloromethane)
to afford title product (300 mg, 67%); MS (AP+) m/e 446
[M+H].sup.+
Step 5.
5-(1-Hydroximino-indan-5-yl)-4-pyridin-4-yl-thiophene-2-carboxyli-
c acid methyl-(1-methyl-piperidin-4-yl)-amide
The product of step 4 (300 mg, 0.8 mmol) was heated at reflux in
ethanol (10 ml) containing hydroxylamine (50% aqueous solution) (1
ml) for 1 hour. The solution was co-evaporated in ethanol
(.times.3) to afford a crude solid. The product was purified twice
by silica gel chromatography (1:9:40 0.880
ammonia:thanol:dichloromethane) to afford the title compound (80
mg, 22%); MS (AP+) m/e 461 [M+H].sup.+
It is to be understood that the present invention covers all
combinations of particular and preferred subgroups described
hereinabove.
BIOLOGICAL EXAMPLES
The activity of compounds of formula (I) as B-Raf inhibitors may be
determined by the following in vitro assay:
Fluorescence Anisotropy Kinase Binding Assay
The kinase enzyme, fluorescent ligand and a variable concentration
of test compound are incubated together to reach thermodynamic
equilibrium under conditions such that in the absence of test
compound the fluorescent ligand is significantly (>50%) enzyme
bound and in the presence of a sufficient concentration
(>10.times.Ki) of a potent inhibitor the anisotropy of the
unbound fluorescent ligand is measurably different from the bound
value. The concentration of kinase enzyme should preferably be
.gtoreq.1.times.K.sub.f. The concentration of fluorescent ligand
required will depend on the instrumentation used, and the
fluorescent and physicochemical properties. The concentration used
must be lower than the concentration of kinase enzyme, and
preferably less than half the kinase enzyme concentration. A
typical protocol is:
All compounds are serially diluted in DMSO, then by a one step
dilution into buffer of comparison, 50 mM HEPES, pharmaceutical
pH7.5, 1 mM CHAPS, 10 mM MgCL.sub.2, for the assay.
B-Raf Enzyme concentration: 1 nM
Fluorescent ligand concentration: 0.5 nM
Test compound concentration: 0.5 nM-100 uM
Components incubated in 10 ul final volume in LJL HE 384 type B
black microtitre plate until equilibrium reached (Over 3 h, up to
30 h)
Fluorescence anisotropy read in an LJL Acquest fluorescence
reader.
Definitions:
Ki=dissociation constant for inhibitor binding Kf=dissociation
constant for fluorescent ligand binding
The fluorescent ligand is the following compound:
##STR00012##
which is derived from
5-[2-(4-aminomethylphenyl)-5-pyridin-4-yl-1H-imidazol-4-yl]-2-chloropheno-
l and rhodamine green.
Compounds of the invention have a K.sub.d of less than 1 .mu.M.
Raf Kinase Assay
Activity of human recombinant B-Raf protein was assessed in vitro
by assay of the incorporation of radiolabelled phosphate to
recombinant MAP kinase kinase (MEK), a known physiologic substrate
of B-Raf. Catalytically active human recombinant B-Raf protein was
obtained by purification from Sf9 insect cells infected with a
human B-Raf recombinant baculovirus expression vector. To ensure
that all substrate phosphorylation resulted from B-Raf activity, a
catalytically inactive form of MEK was utilised. This protein was
purified from bacterial cells expressing mutant inactive MEK as a
fusion protein with glutathione-S-transferase (GST-kdMEK). Method:
Standard assay conditions of B-Raf catalytic activity utilised 3 ug
of GST-kdMEK, 10 uM ATP and 2 uCi .sup.33P-ATP, 50 mM MOPS, 0.1 mM
EDTA, 0.1M sucrose, 10 mM MgCl.sub.2 plus 0.1% dimethylsulphoxide
(containing compound where appropriate) in a total reaction volume
of 30 ul. Reactions were incubated at 25.degree. C. for 90 minutes
and reactions terminated by addition of EDTA to a final
concentration of 50 uM. 10 ul of reaction was spotted to P81
phosphocellulose paper and air dried. Following four washes in ice
cold 10% trichloroacetic acid, 0.5% phosphoric acid, papers were
air dried prior to addition of liquid scintillant and measurement
of radioactvity in a scintillation counter. Results: The compounds
of the examples were found to be effective in inhibiting B-Raf
mediated phosphorylation of GST-kdMEK substrate having IC.sub.50's
of <3 .mu.M. The activity of compounds as Raf inhibitors may
also be determined by the assays described in WO 99/10325;
McDonald, O. B., Chen, W. J., Ellis, B., Hoffman, C., Overton, L.,
Rink, M., Smith, A., Marshall, C. J. and Wood, E. R. (1999) A
scintillation proximity assay for the RafEK/ERK kinase cascade:
high throughput screening and identification of selective enzyme
inhibitors, Anal. Biochem. 268: 318-329 and AACR meeting New
Orleans 1998 Poster 3793.
The neuroprotective properties of B-Raf inhibitors may be
determined by the following in vitro assay:
Neuroprotective Properties of B-Raf Inhibitors in Rat Hippocampal
Slice Cultures
Organotypic cultures provide an intermediate between dissociated
neuronal cell cultures and in-vivo models of oxygen and glucose
deprivation (OGD). The majority of glial-neuronal interactions and
neuronal circuitry are maintained in cultured hippocampal slices,
so facilitating investigation of the patterns of death among
differing cell types in a model that resembles the in vivo
situation. These cultures allow the study of delayed cellular
damage and death 24 hours, or more, post-insult and permit
assessment of the consequences of long-term alterations in culture
conditions. A number of laboratories have reported delayed neuronal
damage in response to OGD in organotypic cultures of the
hippocampus (Vornov et al., Stroke, 1994, 25, 57-465; Newell et
al., Brain Res., 1995, 676, 38-44). Several classes of compounds
have been shown to protect in this model, including EAA antagonists
(Strasser et al., Brain Res., 1995, 687, 167-174), Na channel
blockers (Tasker et al., J. Neurosci., 1992, 12, 98-4308) and Ca
channel blockers (Pringle et al., Stroke, 1996, 7, 2124-2130). To
date, relatively little is known of the roles of intracellular
kinase mediated signalling pathways in neuronal cell death in this
model. Method: Organotypic hippocampal slice cultures were prepared
using the method of Stoppini et al., J. Neurosci. Methods, 1995,
37, 173-182. Briefly, 400 micron sections prepared from hippocampi
of 7-8 day postnatal Sprague Dawley rats are cultured on semiporous
membranes for 9-12 days. OGD is then induced by incubation in serum
and glucose-free medium in an anaerobic chamber for 45 minutes.
Cultures are then returned to the air/CO.sub.2 incubator for 23
hours before analysis. Propidium iodide (PI) is used as an
indicator of cell death. PI is non toxic to neurones and has been
used in many studies to ascertain cell viability. In damaged
neurons PI enters and binds to nucleic acids. Bound PI shows
increased emission at 635 nm when excited at 540 nm. One PI
fluorescence image and one white light image are taken and the
proportion of cell death analysed. The area of region CA1 is
defined from the white light image and superimposed over the PI
image. The PI signal is thresholded and area of PI damage expressed
as a percentage of the CA1 area. Correlation between PI
fluorescence and histologically confirmed cell death has been
validated previously by Niss1-staining using cresyl fast violet
(Newell et al., J. Neurosci., 1995, 15, 7702-7711).
The anti-cancer properties of compounds of the invention may be
determined by the following in vitro assays:
Methylene Blue Growth Inhibition Assay (Assay 2)
Normal human foreskin fibroblasts (HFF), human melanoma (A375P,
SKMEL2, SKMEL3) colon carcinoma (Colo 205) were cultured in the
following growth media: A375P, Colo 205, Roswell Park Memorial
Institute (RPMI) 1640 (Life Technologies 22400-089) containing 10%
fetal bovine serum (FBS); HFF, Dulbecco's modified Eagle Medium
(DMEM) (Life Technologies 12320-032) containing 10% FBS; SKMEL2 and
SKMEL3, Minimum Essential Medium (MEM, Life Technologies 11095-080)
containing 1.times. non-essential amino acids (Life Technologies
11140-050) and 10% FBS. Cells were harvested using 0.25% trypsin/1
mM, EDTA, counted using a haemocytometer, and plated in 10.0
microliters of the appropriate media, at the following densities,
in a 96-well tissue culture plate (Falcon 3075): BFF and A375P,
5,000 cells/well; all other cell lines, 10,000 cells/well. The next
day, compounds were diluted in RPMI containing 100 micrograms/ml
gentamicin, at twice the final required concentration, from 10 mM
stock solutions in dimethyl sulphoxide (DMSO). One hundred
microliters per well of these dilutions were added to the 100
microliters of media currently on the cell plates. RPMI containing
0.6% DMSO was added to control wells. Compounds diluted in. The
final concentration of DMSO in all wells was 0.3%. Cells were
incubated at 37.degree. C., 5% CO.sub.2 for 3 days. Medium was
removed by aspiration. Cell biomass was estimated by staining cells
with 90 .mu.l per well methylene blue (Sigma. M9140, 0.5% in 50:50
ethanol:water) and incubation at room temperature for at least 30
minutes. Stain was removed, the plates rinsed by immersion in
deionized water and airdied. To release stain from the cells 100
.mu.l of solubilization solution was added (1% N-lauroyl sarcosine,
sodium salt, Sigma L5125, in phosphate-buffered saline solution
(PBS)), and plates were incubated at room temperature for 30
minutes. Optical density at 620 nM was measured on a microplate
reader. Percent inhibition of cell growth was calculated relative
to vehicle treated control wells. Concentration of compound that
inhibits 50% of cell growth (IC.sub.50) was interpolated using
nonlinear regression (Levenberg-Marquardt) and the equation,
y=V.sub.max*(1-(x/(K+x)))+Y2, where "K" was equal to the
IC.sub.50.
XTT 72 hr Growth Inhibition Protocol for Mammalian Cultured Cells
(Assay 3)
Human diploid foreskin fibroblasts (HFF) or human colon carcinoma
(Colo 201) cells were grown in Dulbecco's modified Eagle's medium
(DMEM) (Invitrogen/Life Technologies) containing 10% fetal bovine
serum (FBS) and the antibiotics penicillin (100 Units/ml) and
streotomycin (100 micrograms/ml) (Invitrogen/Life Technologies).
Growth was at 37.degree. C. in humidified 5% CO2 incubators in 75
cm.sup.2 plastic flasks. Cells were harvested using 0.25% trypsin/1
mM ethylenediaminetetraacetic acid (EDTA), resuspended in growth
medium, and counted using a hemocytometer. Flat-bottomed 96-well
plates were seeded with 2.times.10.sup.3 cells/well in a volume of
200 ul from trypsinized exponentially growing cultures. To "blank"
wells, growth medium was added with no additions. Cells were
incubated overnight to permit attachment.
Next day, medium from wells that contained cells was replaced with
180 microliters of fresh medium. Appropriate dilutions of test
compounds were added to the wells from stock soloutions of compound
dissolved in dimethyl sulfoxide (DMSO); final DMSO concentration in
all wells was 0.2%. Cells plus compound were incubated for an
additional 72 hr at 37.degree. C. under normal growth conditions.
Cells were then assayed for viability using standard XTT/PMS*.
Fifty microliters of XTT/PMS solution was added to each well and
plates were incubated for 90 minutes at 37.degree. C. Absorbance at
450 nM was then determined using a 96-well UV plate reader
(Molecular Devices). Under these conditions, absorbance of
untreated control cells at 450 nm was at least 1.0 optical density
unit/ml. Percent viability of cells in each well was calculated
from these data (having been corrected for background absorbance).
It was equal to 100.times.(A450 test well/A450 untreated control
well), the A450s being averages of triplicate determinations. IC50
was that concentration of compound that reduced cell viability to
50% of control (untreated) viability, as determined from plots of
concentration vs percent viability. Preparation of XTT/PMS solution
(immediately before assay). For each 96-well plate, 8 mg XTT
(2,3-bis[2-Methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide)
(Sigma Chemical Co.) per plate was dissolved in 100 ul DMSO. 3.9 ml
H.sub.2O was added to dissolve XTT and 20 ul of PMS (phenazine
methosulfate, Sigma Chemical Co.) stock solution (30 mg/ml) was
added from frozen aliquoted stock solution (10 mg of PMS in 3.3 ml
phosphate buffered saline (vitrogen/Life Technologies). (These
stocks are routinely frozen at -20.degree. C. until use). Normal
human foreskin fibroblasts (BFF) are the control normal cell line
that should not be inhibited or at least much less sensitive.
TABLE-US-00006 Cell Line HFF Colo201 Colo205 A375P SKMEL3 SKMEL2
Pathology normal Colorectal cancer Colorectal cancer melanoma
melanoma melanoma B-Raf wt ND V599E V599E V599E wt Status B-Raf,
Ras wt ND wt wt wt [Q61R]N- nM Kd Status Ras Example Assay 1 Assay
2 Assay 3 Assay 2 Assay 2 Assay 2 Assay 2 No 20 0.4 >30*
0.8.sup..DELTA. 0.32.sup..DELTA. 0.31.sup..DELTA. 0.46.sup-
..DELTA. 1.9.sup..DELTA. *indicates IC50 >3 .mu.M
.sup..DELTA.indicates IC 50 0.3-3 .mu.M .sup..dagger.indicates IC50
<0.3 .mu.M A375, Colo205 and SKMEL are reported as wild type
(wt) for Ras status in the literature. V599E indicates that the
cell lines have activating BRaf mutation (V599E) ND represents not
determined
Throughout the specification and the claims which follow, unless
the context requires otherwise, the word `comprise`, and variations
such as `comprises` and `comprising`, will be understood to imply
the inclusion of a stated integer or step or group of integers but
not to the exclusion of any other integer or step or group of
integers or steps.
The application of which this description and claims forms part may
be used as a basis for priority in respect of any subsequent
application. The claims of such subsequent application may be
directed to any feature or combination of features described
herein. They may take the form of composition, process, or use
claims and may include by way of example and without limitation the
following claims.
* * * * *